SU1386656A1 - Strain consortium of leuconostoc oenos bacteria used for decreasing sourness of grape must and wine products - Google Patents

Abstract

The invention relates to biotechnology and relates to an associative culture consisting of strains of lactic acid bacteria of the species Leuconostoc olnos, which can be used in winemaking culture. The aim of the invention is to obtain a new consortium of bacterial strains Leuconostoc olnos VKSh B-3365 (Tairovska 1), which has a higher biological activity of acid degradation in the wort and in the wine material of different varieties and of different composition and capable, in turn, to dispose of malic acid at a low value pH, high content of tartaric acid, sulfuric anhydride and at low temperatures. The new consortium contains the following bacterial strains: T-A12, T-118, T-120, T-2C and T-96 in a ratio of 20:20:20: 20: 20%. These strains were obtained as a result of the selection of production microorganisms and isolated from dry Belk wine materials of Aligote and Rkatsiteli, the Consortium is able to carry out the process of block-milk fermentation in Aligote wine material with a content of titratable acids of 13.3 g / l, tartaric acid of 7.8 g / l, sulfur dioxide 10.0 mg / l free and 70.0 mg / l total, at pH 2.75 for 14 days. The titration of 1x acids in the process of biological acid lowering decreases from 11.6 to 6.3 g / l. At the task of the consortium of lactic acid bacteria Tairovsk -1, the non-material-fermented fermentation fermentation has a low-grade varietal aroma, soft in taste and received a tasting score of 8.1–8.3 points. 8 tab. I (L with 00 O) O) 01 O5

Description

This invention relates to biotechnology and relates to an associative culture consisting of strains of lactic acid bacteria of the species Leuconosto olnos, which can be used in the wine industry.

The purpose of the invention is to obtain a new consortium of lactic acid bacteria of the species Leuconostoc olnos, which possesses a higher activity of biological acid lowering in wort and wine materials of different varieties and of different composition and ability

dispose of malic acid at low pH, high tartaric acid, sulfuric anhydride and gfI low temperatures

The bacteria consortium Leuconostoc olnos BKliM B-3365 (Tairovska -1) contains the following strains of lactic acid bacteria: T-A, T-118; T-120 T-2C and T-96 in the ratio .20: 20: 20:: 20; 20%

The strains belonging to the consortium were isolated from white varieties of wine materials: T – A – T – 2C and T – 96 –– from the grape material Aligote 22 Nvar

1982 .. T-120 and T-118 - from Ramaciteli 30 brand wine brand

1983

The first separation of bacteria was carried out according to the following scheme:

sowing of 1% of the wine material being studied on medium of MPC + wine (1: 1) + levorin + + sing 1/2 vaseline oil;

reseeding after 48 h on solid culture medium MPC, incubation at t 28 ° C;

isolation of lactic acid bacteria from individual colonies.

The strains are characterized by the following morphological, cultural, and physiological and biochemical characteristics.

Morphological and cultural signs.

All strains included in the consortium (T – A, g, T – 118, T – 120, T – 2C and T – 96) are Gram positive fixed cocci of 0.7–1.0 µm, facultative anaerobes. On an agar medium, MPC at t 28 C on the second day give a weak growth in the form of white smooth translucent colonies with a diameter of 0.1

1 mm. In liquid medium, MPC + wine (1; 1) forms whitish dregs with a pronounced sediment, which rises up in the form of silky waves when agitated. Do not form mucus.

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Physiological and biochemical signs.

Strain T-A forms a gas during the decomposition of glucose and does not form ammonia from arginine. Ferments glucose, arabinose, maltose, sucrose, cellobiose; does not ferment xylose, lactose, melibiosis, raffinose, mannnt, sorbitol, glycerin. Does not restore nitrates and nitrites, does not liquefy zhelmatin. Does not absorb citrate, absorbs malate, does not decompose tartrate. Catal azo-negative. The strain T-118 forms a gas during the decomposition of glucose, does not form ammonia from arginine. Ferments arabinose, xylose, sucrose, maltose, melibiosis, sorbitol; does not ferment lactose, galactose, raffinose, maltose, cellobiose. Does not restore nitrates to nitrites, does not dilute gelatin. Does not absorb citrate And tartrate. It is absorbed by malate, catalase-negative,

The strain forms a gas during the decomposition of glucose, does not form ammonia from arginine. It ferments glucose, arabinose, mannose, maltose, cellobiose, does not ferment galactose, lactose, raffinose, sucrose, mannitol, sorbitol. Does not restore nitrates to nitrite, does not dilute gelatin, does not decompose glycerin. It does not absorb citrate and tartrate, decomposes malate, catalo-negative.

Strain T-120 with the decomposition of glucose forms a gas that does not form ammonia from arginine. Ferments glucose, arabinose, xylose, mannose, galactose, cellobiose, does not ferment lactose, raffinose, mannitol, sorbitol. Does not restore nitrates to nitrites, does not dilute gelatin, does not decompose glycerin. Does not absorb citrate and tartrate. Absorbs malate, catapas-negative.

The strain T-96 in the decomposition of glucose forms a gas, does not form ammonia from arginine. Ferments glucose, arabinose, xylose, maltose, sucrose, does not ferment galactose, mannose, cellobiose, raffinose, mannitol. Does not restore nitrates to nitrites, does not dilute gelatin, does not decompose glycerin. Does not absorb citrate and tartrate, decomposes malate. Catalase-negative

Some strains of lactic acid bacteria can accumulate in wines unwanted nutrient amine - his15

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tamin, which significantly impairs the taste of these wines.

All strains of lactic acid bacteria that are part of the associative

Tairovska cultures -1, do not form histamine from histidine,

 The consortium of strains is stored in a liquid medium, consisting of MPC and vii. Material 1: 1, with re-sowing once every 3 months. At the same time, the culture retains a high biochemical activity.

The composition of the medium MLS:

Peptone10g

Yeast

autolysate 50 ml

M sleep water 100 ml

Twin-801 ml

Glucose 20 g

Acetic acid

sodium 5 g

Citric acid

ammonium triehame

schenny () 2 g

Phosphate

potassium dibasic

(KjHP04-3HiO) 2 g

Sulphate

magnesium

(MgS04-7HiO) 0.2 g

Sulfuric margar

hell

(nSOv4H-lO) 0.05 g

Distilled

waterUp to I l

The consortium of VKPM B-3365 strains was obtained by co-cultivation in a MES + wine medium (1: J) lactic acid bacteria — strain T-A, T-120, T-P8, T-2C T-96. The culture medium was inoculated with the indicated strains simultaneously in equal ratios (1: 1: 1: 1: 1), -Bacteria were grown for 30 days with weekly replanting

3g

40

45

25

thirty

The selection of the best strains from 120 isolated cultures of lactic acid bacteria in comparison with production analogs was carried out taking into account the requirements imposed on microorganisms using 1M for acid-reduction of grape must and table wine materials, Chmumml should have high acid resistance, sulfite resistance, cold resistance, sulfite resistance, cold resistance, sulfite resistance, cold resistance, sulfite resistance, cold resistance. ,

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The ability of bacteria to utilize maternal acid, with different contents of titratable acids, tartaric acid and pH, was determined on the alumina variety vitamins in five variants with free sulfuric anhydride content of 8, 10 and 11 mg / l, total 60, 70 and 75 mg / l (table 1).

The distribution of lactic acid bacteria was introduced in the amount of 5% into wine material, poured into 50 ml flasks. The acidic capacity of lactic acid bacteria was tested according to the following indicators: content of titratable acids, pH, qualitative determination of block and lactic acids by paper chromatography.

Bacteric acid utilization of bacteria was ascertained by a decrease in titratable acidity, the disappearance of malic acid, an increase in pH, and the rate of block-lactic fermentation (NMB) was also taken into account.

The properties of the most active strains are listed in tabLo2,

From the data of Table 2 it can be seen that strains T-118, T-120, T-96, T-A „are capable of utilizing block acid in wine materials at pH 2.8. Strains T-A., And T-2C dispose of maternal acid at a content of free sulfuric anhydride of 11.0 mg / l. After obtaining a consortium from

The g strains were re-tested for the acid-lowering ability of each strain included in the association and associative culture (Table 3).

The test was carried out on a wine-making grade Apigote. As can be seen from Table 3, an associative culture is capable of utilizing malic acid in a wine material with a titrated acidity of 13.3 g / l and a tartaric acid content

5 7 g / l, sulfurous anhydride free 11 mg / l and total 75 mg / l; pH 2.80 for 14 days, active strain5

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we are T-A and T-120 for 30 days. The strains T-2C, T-96 and T-118 only partially block-milk fermentation under extreme conditions.

The acid-lowering ability of the consortium was tested in high-acid wine material of the variety Aligote with various combinations of block and tartaric acids (Table 4).

The data from the scoreboard4 indicates that if the acidity is titrated

represented mainly by tartaric acid, the associative culture is able to dispose of malic acid: in the wine material with a titanium acid content of 13 g / l (the amount of j trescent is 7.4 g / l). When large amounts of malic acid are contained in the wine material (the amount of tartaric acid is 3.4 g / l), the culture conducts the process of block-lactic fermentation with a titrated acidity of 15 g / l.

The consortium and its strains were tested for acid lowering ability at 10, 18 and 25 Ce 15 material is more stable.

13866566

acids (0.1 g / l more than the control). In the experimental wine materials, recovery processes are well performed. The redox potential decreased from 409 to 364 mV against 384 mV in the control sample, the amount of aldehydes decreased, and the content of reductants increased.

The content of nitrogenous substances is lower Also in the experimental wine material, where the block-milk fermentation took place

completely. Consequently, this wine was studied on sterile wine Rkatsiteli of the following chemical composition; co.- the titration of titratable acids 10.8 g / l, tartaric acid g / l, pH 3.2, the content of sulfurous anhydride free 10 mg / l and the total 60 mg / l (table 5).

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less prone to cloudiness and over oxidation. This wine sample was soft, harmonious, full in taste, had a well-blended varietal aroma and received a high tasting rating of 8.3 points. The control sample of wine materials received a score of 7.9 points due to rough green acidity, inharmony in the taste of

As can be seen from the scoreboard5, the resulting consortium of strains utilizes the block sump at t 25 ° C for 5 days, at t 8 ° C for 12 days, at t 10 ° C for 121 days. The strains T-Ac, T-96 and T-2C performed the block-milk process, fermentation in 7 days, at t 18 С in 114 days, at t 10 ° С for 21 days. Strains T-120 and T-118 acid reduction is carried out at 25 ° С for 10 days, at t 18 С for 21 days, at t 10 ° С for 25 days

Table 6 presents a comparative characteristic of white table Wine of Apigote variety, prepared for semi-production conditions with biological acid reduction using the Tairovsk-1 culture. Time25

thirty

35

40

less prone to cloudiness and oxidation pen. This sample of wine rial was soft, harmonious, in taste, had a rich varietal aroma, and received a high tasting score of 8.3 points. The control sample of wine materials received a score of 7.9 points due to rough green acidity, inharmony in the taste of

Comparative characteristics of sortium strains of Tairovsk -1 and K-48F are presented in Table 7. ,

Example 1. A strain consortium is grown in grape must with ta Cabernet diluted 1: 1. The obtained 2-day wiring of the batteries in the amount of 3-5% is set in the capacity V - 160 decalit. Wine material has the following conditions: containing titratable acids 10 g / l, tartaric acid 4 g / l, pH 3.1, content of blue anhydride free 15 m and total 75 mg / l After 5 the acidity of the test wine cellar decreases to 5, 6 g / l, pH increases to 3.5, according to chromatographic studies, the total disappearance

Bacteria were prepared in lab-dacidic acid. Wine after wine conditions on grape mash, acid reduction is mild,

harmonious

| Diluted with water in a ratio of 1: 1, and 160 l were introduced into a container with V-material.

The tablob data testify to troM that the culture is capable of carrying out .t-modo-block-milk fermentation. In wine materials with high initial (titrated acidity (13.2 g / l) 1c high content of tartaric acid (7.0 g / l). Titrated acidity .Yri this decreased from 13.2 to 7.8 g / l iipH slight accumulation of volatile

tastes and has a valuation score of 8.2 points in comparison with a control sample, which has a score of 7.9 points.

50Example2. Bacterial p

Culture vodka is accumulated in cultivators filled with 2/3 vol. of pasteurized wort, and 5–10% of the wiring is set. . Cultivation pier

55 acidic bacteria are carried out at a constant temperature and occasional stirring for an equally (; - Mercury distribution

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less prone to cloudiness and over oxidation. This wine sample was soft, harmonious, full in taste, had a well-blended varietal aroma and received a high tasting rating of 8.3 points. The control sample of wine materials received a score of 7.9 points due to rough green acidity, inharmony in the taste of

The comparative characteristics of the consortium of Tairovsk-1 and K-48F strains are presented in Table 7. ,

Example 1. A consortium of strains is grown in grape must Cabernet diluted 1: 1. The resulting 2-day wiring of bacteria in the amount of 3-5% set in the capacity of V - 160 gave. Wine material i has the following conditions: the content of titratable acids is 10 g / l, tartaric acid is 4 g / l, pH 3.1, the content of sulfuric anhydride is free 15 mg / l and total 75 mg / l After 5 days, the acidity of the wine stock studied is reduced to 5 , 6 g / l, pH rises to 3.5, according to chromatographic studies, the complete disappearance of the harmonic

in taste and has a tasting score of 8.2 points compared with the control sample, which has a score of 7.9 points.

Example2. Bacterial wiring culture accumulate in the cultivators, filled with 2/3 of the volume of pasteurized wort, set 5 - 10% wiring. The cultivation of lactic acid bacteria is carried out at a constant temperature and occasional stirring for equal (; - Measured distribution of the bacterial mass.

When the cultivator has a titer of 10-10 kg / MP in the entire volume of the cultivator, the wiring is used for biological acid lowering, pumped in a tank V 1250 with high acid wort and wine material in an amount of 5% by volume. when 1 2-20 C, the Results are presented in table 8.

The wine materials in which the block-milk fermentation took place at the task of the consortium of lactic acid bacteria Tairovsk t, had a pronounced varietal aroma, were soft in taste and received a high tasting rating.

Claims (1)

Invention Formula -  Bacterial Strain Consortium Leuconostcc olnos VKPM B-3365, isp acid lowering wine and wine materials. Table 1 8.0 / 60.0 11.0 / 75.0 8.0 / 6Q, 0 11.0 / 75.0 8.0 / 60.0 7.8 4.5 4.5 4.5 7.8 13.3 8.6 8.6 8.6 13.3 Consort tsuk 11/75 table 2 2.8 3.2 3.2 3.2 2.8 3 3.33 3.33 3.33 3.1 thirty 14 7 14 thirty t a b l  10.4 2.8 3.1 +++ ++ 4 ++ H-1- + +++ . ++ 30 30 30 30 30 14  Table Original wort 3.4 489 "3, L 7.2 0.5 + + and output number 2.9 after x lottery baggage with Taic POBCXMI 3.3 culture 40913.2 7.0 0.2 ++ ++ 364 7.8 6.5 0.3 +++ it is source 19.8 80.0 10.8 186 after acidoilicon with culture Tshgroisk-1 15.275.0 9.5 170 Table 368 185 336 174. + 274 UO Continued, table. 191 v6.0 11.2 12 7.9 180 80 11.5 P 8.3 2 2 I'm to I am  l 8 I 2 8 s s And s 28 about IS  gh - oh oh J i n I II I .1 111 I I I i 4D m "M m  to Iff "h (n "" N "" (4 "h “H“ o M / y 4 o “o " s about " " - % N (I'm about oh :: "0" in rt in about  l f "In in